We are sorry, but NCBI web applications do not support your browser and may not function properly. More information

Results: 5

1.
Fig. 5

Fig. 5. From: Molecular basis for the high degree of antigenic cross reactivity between hepatitis B virus capsids (HBcAg) and subunits: Insights into the enigmatic nature of e-antigen.

Characterization of Cp149d:Fab e6 and Cp(-10)149d:Fab e6 immune complexes. Analysis of the complexes by (a) gel filtration chromatography, (b) analytical ultracentrifugation, and (c) reducing SDS-PAGE indicates a 1:2 molar ratio in both cases.

Norman R. Watts, et al. J Mol Biol. ;398(4):530-541.
2.
Fig. 3

Fig. 3. From: Molecular basis for the high degree of antigenic cross reactivity between hepatitis B virus capsids (HBcAg) and subunits: Insights into the enigmatic nature of e-antigen.

Typical sensograms illustrating the binding of antigen to Mab 3105. Antigen was allowed to bind to immobilized antibody. Duplicate, and in some cases triplicate, injections of antigen were done. The concentrations refer to the concentration of antigen, in terms of dimers. The kinetics and affinities of Cp149c (A) and Cp149d (B) for Mab 3105 are similar, indicating that the epitope is not affected significantly by the assembly state of the antigen. This is in keeping with its known location on the apex of the spike(s).

Norman R. Watts, et al. J Mol Biol. ;398(4):530-541.
3.
Fig. 4

Fig. 4. From: Molecular basis for the high degree of antigenic cross reactivity between hepatitis B virus capsids (HBcAg) and subunits: Insights into the enigmatic nature of e-antigen.

Immune complexes visualized by electron microscopy. Cp(-10)149d were mixed with antibodies in solution and visualized in negative stain. (A) Mab alone, (B) Dimer + Mab 904, (C) Dimer + Mab 905. Outlines and an interpretative diagram are shown in each case. The formation of the circular complexes in (C) suggests the presence of two copies of the 905 epitope per Cp(-10)149d molecule. Very similar complexes were observed when Mab e6 was employed (not shown). Bar = 200 Å.

Norman R. Watts, et al. J Mol Biol. ;398(4):530-541.
4.
Fig. 2

Fig. 2. From: Molecular basis for the high degree of antigenic cross reactivity between hepatitis B virus capsids (HBcAg) and subunits: Insights into the enigmatic nature of e-antigen.

Locations of the epitopes on HBV capsids and subunits. The cartoon represents three adjacent dimeric subunits on a capsid with the protruding spikes corresponding to the four-helix bundles. The epitopes for Mabs 88, 312, 842, 3105, 9c8, and F11A4 are all located on the apices of the spikes whereas the epitope for Mab 3120 is located between the spikes and involves residues from two adjacent subunits.23; 24; 25; 26; 27 The two copies of the Mab e6 epitope are located near the C-termini of the dimer and are accessible only on free subunits. The locations of the epitopes for Mabs 904 and 905 are uncertain, but epitope 904 may be on the spike apex and 905 near the C-terminus (see text). The positions of the three carboxy-terminal mutations employed to map the e6 epitope, and which affect capsid assembly, are also indicated.

Norman R. Watts, et al. J Mol Biol. ;398(4):530-541.
5.
Fig. 1

Fig. 1. From: Molecular basis for the high degree of antigenic cross reactivity between hepatitis B virus capsids (HBcAg) and subunits: Insights into the enigmatic nature of e-antigen.

Schematic representation of the recombinant HBV proteins used in this study. The three capsid-related proteins (Cp) correspond to those listed in Table 1 and in greater detail in Fig. 2. Cp183 is expressed in E. coli as nucleic acid-filled capsids (Cp183c) with a mass of ∼6 MDa39 (represented by the blue fenestrated structure). Such capsids are very stable and cannot be dissociated into subunits without protein denaturation. In Cp149 the arginine rich carboxy-terminal domain has been deleted and when this protein is expressed in E. coli it forms empty capsids (Cp149c) with masses of 3 and 4 MDa39 (represented by a grey fenestrated structure). These capsids can be reversibly dissociated into dimeric subunits with a mass of ∼35 kDa (grey object). The red line indicates an intermolecular disulfide bond [C61-C61]. The Cp(-10)149 or e-antigen is expressed in E. coli in a form that is neither assembled capsid nor an aggregated inclusion body-type protein. The protein can be extracted with 2 - 3 M urea at pH 9.5 to give soluble folded dimers. Purified dimers can be induced to form capsids (white fenestrated structure). The reversibility of this has not been fully explored. The red lines indicate intramolecular disulfide bonds [C(-7) – C61]. See Figs. S1 and S2 for analytical data on these various proteins.

Norman R. Watts, et al. J Mol Biol. ;398(4):530-541.

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk